The overarching theme of this supplementary project is to add the new information of parent award that nutrition lead to the changes of extracellular vesicles (EVs) of IECs in alcoholic liver disease (ALD). We will focus on ?6 PUFAs and gut microbiota-regulated exmiRNAs. Specifically, we will examine how ?6 PUFAs stimulate the production of gut EVs through TLR4/STAT3 signaling and the consequences of these events in the development of ALD via gut-liver axis. Novel in vitro and in vivo approaches will be used to address mechanisms of action with the goal of identifying new targets for intervention. We propose that ?6 PUFAs induce the dysbiosis of gut microbiota, which contribute to the changes in the miRNAs profile of gut-derived EVs. We specifically propose that TLR4/STAT3-activated extracelluar miRNAs production is the key to the activation of macrophages during ?6 PUFAs-induced chronic inflammation in ALD. Based on our published work and other preliminary data, this proposal will test the hypothesis that ?6 PUFAs regulate the production of gut extracellular miRNAs through TLR4/STAT3 signaling, which initiates a crosstalk among macrophages and hepatocytes in ALD. Moreover, we will determine whether supplement of ?3 PUFAs with diet-associated nanoparticles carrying inflammatory miRNAs inhibitor have potential therapeutic implications in ALD. These hypotheses will be tested in the following Specific Aims: Aim 1: Determine the role of ?6 PUFAs-regulated gut EVs in alcohol-induced liver inflammation. Aim 2: Determine whether alteration of IEC miRNAs via TLR4/STAT3 modulation affects the severity of ?6 PUFAs-induced liver inflammation and injury in ALD. .